During early outgrowth of Bacillus subtilis spores, there is a period of RNA and protein synthesis in the absence of DNA replication. Two mutants of B. subtilis, PB2442 (gsp-4) and PB2452 (gsp-81), were used to study the pattern of RNA synthesis during this period. The two mutants are temperature-sensitive in the outgrowth phase, and show a limited amount of incorporation of [3H]uridine at 47 °C. The RNAs synthesized during a 2 min pulse with [3H]uridine were hybridized to Eco RI-digested DNA, after agarose gel electrophoresis and transfer to nitrocellulose paper (Southern technique). For both mutants the transcripts synthesized at 35 °C at different times were different. Differences were also observed in the transcripts made at 47 °C. For both mutants, in the presence of chloramphenicol, the same hybridization pattern was obtained for RNAs pulse-labelled at different periods during outgrowth.
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